NCL Method ITA-3
Mouse Granulocyte-Macrophage Colony-Forming Unit Assay
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Version 2.0 - published on 09 Jul 2020 doi:10.17917/0ZHD-SA93 - cite this Last public release: 2.0b
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Description
Myelosuppression is a common dose limiting toxicity of cytotoxic oncology drugs. Nanoparticles may distribute to bone marrow and/or release drug that is delivered to bone marrow. Therefore, understanding potential toxicity of nanoparticles or drugs which nanoparticles carry is an important step in preclinical safety evaluation.
Hematopoietic stem cells of bone marrow (BM) proliferate and differentiate to form discrete cell clusters or colonies. This document describes a protocol for quantitative analysis of granulocyte-macrophage (GM) colony-forming units (CFU), employing murine BM. This protocol can be used for both in vitro and ex vivo analyses. The in vitro protocol involves isolation of bone marrow cells from healthy animals, followed by treatment in vitro with nanoparticle formulations. In the ex vivo version, the bone marrow is isolated from animals injected with the nanoparticle formulation. The in vitro protocol does not account for nanoparticle biodistribution; however, in cases when dose information is not available and nanoparticle formulation is in the early phase of development, the in vitro protocol allows for rapid screening of potentially toxic nanoparticle formulations. The in vitro protocol can also give a quick estimation of the myelosuppressive potential of a cytotoxic oncology drug bound to a nanoparticle surface in comparison to a traditional formulation of the same cytotoxic oncology drug (for an example, see Figure 1).
Content List
- NCL_Method_ITA-3_May2020.pdf(PDF | 695 KB)
Cite this work
Researchers should cite this work as follows:
- Barry Neun, Edward Cedrone, Marina Dobrovolskaia (2020). NCL Method ITA-3. (Version 2.0). NCI Hub. doi:10.17917/0ZHD-SA93
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NCL Protocols
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